Currently, the Japanese Association of Zoos and Aquariums has four roles (species conservation, education and environmental education, survey and research, and recreation). Aquarists have many other tasks to perform including breeding work, planning special exhibitions, events, reception work, sales activities, and product development; thus, they may have limited opportunities to conduct survey and research work. Depending on the size of the aquarium and its staff, the burden on each individual may be high. The difficulty of conducting research in such a situation depends on the target species and the content of the research. In this paper, I introduce some examples of my work, including the discovery of a new species through fieldwork, in the hope that my experiences will be helpful to those who will conduct survey and research in the future.

　In recent years, research activities in zoos have been increasing. This paper describes a report of blood samples taken from a Hartmann's mountain zebra (Equus zebra hartmannae) using husbandry training and its clinical examination data, and a case report on an American beaver (Castor Canadensis). These reports were based on data collected by daily routine work at Hitachi Kamine Zoo. It is important to collect and store data, which may lead to publishing these kinds of reports. I hope that this paper will provide readers with insights into their research activities.

　In recent years, research has become increasingly popular in zoos. However, zoos are often faced with the following three problems in efforts to conduct research: (1) insufficient knowledge, (2) budgetary constraints, and (3) insufficient time available. We explored methods for solving these three problems by examining the research efforts of the Chiba Zoological Park. There are three priorities that need to be addressed: (1) Accumulation of experience by hiring dedicated research staff and building a track record of research programs with successful results at the zoo. (2) Obtaining research funding from the city and from various additional sources such as grants and donations. (3) Accumulating a small number of samples over longer timescales. Methods of behavior evaluation can then be undertaken by accumulating a small number of samples over a relatively long period of time, with the aim of collaborating with “family-oriented research institutes.”

　In recent years, research and educational activities at zoos have become increasingly common. Nevertheless, the perception of zoos as research and educational institutes is still not common. Universities, by contrast, have few research results that are widely available to the general public. A strong need exists to conduct more practical research related with local communities and to give back the research results to society. Our university members from the faculties of agriculture, engineering, and education aim to create a new partnership system enhancing the research and educational functions of all collaborators, with local universities supporting local zoos as primary research institutes, and collaborative activities with Hitachi City Kamine Zoo from 2015 and the Chiba City Zoological Park from 2020. For these collaborations, university staff and students cooperate in promoting research using zoos, supporting research activities by zookeepers, and arranging zoo events, etc. Zoos cooperate with university classes and internships. Especially, we assign great importance to the creation of relationships by which staff members can mutually meet face-to-face and feel free to communicate with each other during activities. Collaborations such as our project have already been carried out between other universities and zoos in the past. They are not particularly novel. However, we hope that our activities will serve as a model for local universities and local zoos to become regional academic and cultural hubs through collaborative research and education activities.

　 The objective of this study was to determine the reproductive physiology of captive chamois (Rupicapra rupicapra), including the breeding season and estrous cycle, based on fecal steroid hormone dynamics. We collected fecal samples from two male and one female chamois and quantified fecal testosterone (T) and androstenedione (AD) concentrations in males and progesterone (P4), estradiol-17β (E2), estrone (E1), and cortisol concentrations in the female using enzyme immunoassays. Fecal T and AD concentrations in males showed similar annual variations. Fecal T and AD concentrations in males started to increase in April or May and decreased in November or December when the female estrous cycle resumed. Male to female following, mounting, and copulation were observed during periods of low P4 concentrations in the estrous cycle. The length of the estrous cycle based on P4 dynamics was an average of 23.0 days. The female was already pregnant at the beginning of the present study (79 days before parturition). Fecal P4 concentrations remained high and markedly decreased on the day before parturition. The estrous cycle recurred 195 days after parturition. Fecal E2 and E1 concentrations during pregnancy showed similar high fluctuations. Fecal cortisol concentrations during pregnancy markedly increased approximately 40 days before parturition, remained high until the day before parturition, and then immediately decreased on the day after parturition. The present results revealed that the chamois is polyestrous short-day seasonal breeding animal based on fecal sex steroid dynamics. Furthermore, the measurement of fecal P4, E2, and E1 concentrations in females is useful for the diagnosis and monitoring of pregnancy.

　Asian black bears (Ursus thibetanus) in Japan are clustered into 3 lineages (eastern, western, and southern clusters) based on mitochondrial DNA (mtDNA) haplotypes of an approximately 700 bp sequence in the D-loop region of mtDNA. In 2012 the Ministry of the Environment of Japan announced the Asian black bear population in Kyushu, the southwestern island of the Japanese archipelago, as locally extinct, because the last reliable record of the Asian black bear in Kyushu was in 1941. The number of samples and genetic analysis results pertaining to the Asian black bear population in Kyushu are very few. Therefore, the molecular phylogenetic characteristics of Asian black bears in Kyushu remain a missing link in understanding the trajectory of Asian black bear populations from the Asian mainland to Japan. In this study, we investigated the mtDNA haplotypes of Asian black bears in Kyushu using four old bone samples, which we found at various locations and dates. We succeeded in detecting two new haplotypes and a haplotype of the western cluster. The two new haplotypes also belonged to the western cluster. These results suggest a continuous distribution of Asian black bears between western Chugoku and Kyushu, and indicate that genetic variation occurred after habitat fragmentation. These results provide data to support the considerations of previous studies on the migration of Asian black bears from the Asian continent to the Japanese archipelago.

　Antimicrobial resistance (AMR) in pathogens has become a social problem across humans and wild animals. In particular, the environmental contamination caused by AMR should be monitored. In this study, we analyzed the prevalence of Escherichia coli and its AMR, as well as the distribution and genotype of E. coli in environmental soil and water samples from the habitat of the Hypotaenidia okinawae (Okinawa rail), to elucidate the potential of AMR pollution between the bird and its habitat. The habitat of Okinawa rail was divided into a livestock farm area (LA) near human settlements and a forest area (FA) as a non-human living environment. We found that the prevalence of E. coli in environmental samples collected from the LA was 38.2% (13/34), of which 84.6% (11/13) were antimicrobial resistant. In contrast, the prevalence of E. coli in environmental samples collected from the FA was 25.5% (12/47), of which 16.7% (2/12) were antimicrobial resistant. These results indicate that antimicrobial resistant E. coli was more prevalent in the LA than FA. Furthermore, pulsed-field gel electrophoresis analysis revealed a similar pattern of E. coli prevalence. This suggests that the occurrence of environmental pollution of AMR from livestock farms and that the Okinawa rail may become a carrier of antimicrobial-resistant E. coli in the Yambaru region of Okinawa.

　 The nutria (Myocastor coypus), also known as coypu, is a semi-aquatic rodent introduced to western Japan originally from South America. Knowledge of nutria genetics is needed to elucidate population structure and reproductive biology. This study determined if 27 nuclear microsatellite markers, developed based on nutria introduced to North America, could be used in invasive nutria in Japan. Twenty-two individuals, including four parents and 18 offspring, were used. Amplification success observed at 25 of the 27 loci, which indicated that the marker could be used for genetic analyses of nutria distributed in Japan..

　Female Asian small-clawed otters (Aonyx cinerea) in aquariums and zoos may require sterilization to prevent reproduction and disease or for pedigree-based genetic management. Contraceptive surgery requires ovariohysterectomy or ovariectomy via laparotomy. However, there is a concern that otters may develop various complications due to self-injury at incision sites, especially after procedures requiring longer incisions. Laparoscopic ovariohysterectomy was chosen to prevent these complications in two female otters at the Osaka aquarium KAIYUKAN. To perform laparoscopic ovariohysterectomy, a three-portal technique was used. The surgical durations were 45 and 23 minutes. The minimally invasive procedure was feasible in both cases, and there were no significant complications. Laparoscopic surgery appears suitable for ovariohysterectomy in captive Asian small-clawed otters.